Bovine tuberculosis (bTB) is an endemic zoonosis significantly affects animal health in Burkina Faso. The primary causative agent is Mycobacterium tuberculosis (M. tuberculosis) complex, mainly M. bovis. Cattle are co...Bovine tuberculosis (bTB) is an endemic zoonosis significantly affects animal health in Burkina Faso. The primary causative agent is Mycobacterium tuberculosis (M. tuberculosis) complex, mainly M. bovis. Cattle are considered as natural reservoir of M. bovis. However, in Burkina Faso, the circulation of these strains remains poorly understood and documented. This study aimed to identify and characterize Mycobacterium strains from suspected carcasses during routine meat inspection at Bobo-Dioulasso refrigerated slaughterhouse. A prospective cross-sectional study was conducted from January 2021 to December 2022 on cases of seizures linked to suspected bovine tuberculosis. Microbiological and molecular analyzes were used for mycobacterial strain isolation and characterization. Out of 50 samples, 24% tested positive by microscopy and 12% by culture. Molecular analysis identified 6 strains of Mycobacteria, exclusively Mycobacterium bovis specifically the subspecies bovis (Mycobacterium bovis subsp bovis). In conclusion, M. bovis subsp bovis is the primary agent responsible for bovine tuberculosis in Bobo-Dioulasso. Continuous monitoring of mycobacterial strains is therefore necessary for the effective control of this pathology in the local cattle population.展开更多
The current study was performed to evaluate the liver function status as well as molecular characterization of the recovered worms in rats experimentally infected with F. hepatica. Sixteen male Wister rats aged 30 day...The current study was performed to evaluate the liver function status as well as molecular characterization of the recovered worms in rats experimentally infected with F. hepatica. Sixteen male Wister rats aged 30 days were randomly allocated into two groups (n = 8). The first group was infected orally with 15 viable encysted metacercaria of F. hepatica per animal. The other group was kept non-infected (control group). At zero time (before infection), the 2<sup>nd</sup>, 4<sup>th</sup>, 6<sup>th</sup>, 8<sup>th</sup>, 10<sup>th</sup>, 12<sup>th</sup> and 14<sup>th</sup> weeks post-infection (WPI), blood and serum samples were collected via puncture of retro-orbital plexus of veins from each rat. Serum enzyme level (AST and ALT) and total protein were measured, and the serum protein profile was carried out using agarose gel electrophoresis. During the period of the experiment, serum ALT and AST activities and serum total globulins significantly increased while serum total proteins and albumin markedly decreased in the infected group. On the 14<sup>th</sup> WPI, the data of the electropherogram showed that globulin fractions (α1-, β- and γ-globulin) levels were significantly increased while α2-globulin was markedly decreased in the infected group. The molecular analysis confirmed the amplification of the ITS1, ITS2 and NDI genes of F. hepatica recovered from the infected liver of rats with amplicon sizes of 630, 510 and 560 bp, respectively. Sequencing of the amplified ITS gene resulted in the determination of 3 strains (PP108836, PP108837, and PP108838). Also, analysis of the ITS2 gene resulted in obtaining 3 isolates under the accession numbers (PP109065, PP109066, and PP109067). In conclusion, fasciolosis in the rat model is suitable for routine experimental infections and caused a pronounced liver dysfunction with discharging of the Fasciola eggs in the faeces and the development of adult stages in the bile ducts. Furthermore, molecular techniques are a sensitive tool for the identification and characterisation of the Fasciola parasite.展开更多
Sesame is Burkina Faso’s second essential agricultural export after cotton. It’s consequently a supply of income for producers and foreign exchange for the country. However, sesame production is characterized by low...Sesame is Burkina Faso’s second essential agricultural export after cotton. It’s consequently a supply of income for producers and foreign exchange for the country. However, sesame production is characterized by low average yields of about 538 kg·ha<sup>-1</sup> at the farmer’s field as compared to the potential yield of the improved varieties (1500 - 2000 kg·ha<sup>-1</sup>). Fungal diseases are some of the major constraints to sesame production in Burkina Faso. The present study contributes to the development of means to control pathogenic fungi of this crop, which are responsible for significant losses. The objective is to identify the fungi associated with diseased sesame plant samples. To this end, 149 samples of diseased sesame plants were collected from different production sites located in three agro-climatic zones of the country. The analysis of the samples according to the blotting paper method, based on the morphological characteristics of the fungi, allowed the identification of 18 genera with prevalence rates from 2.68% to 97.98%. The most frequently identified genera were Macrophomina (97.98%), Cercospora (86.57%), Fusarium (85.23%), Phoma (62.41%) and Colletotrichum (61.07%). The results also showed a variable distribution of fungi according to the agro-climatic zone with the predominance of Macrophomina in all three zones. Molecular identification by DNA sequencing of 120 isolates belonging to the different fungi detected allowed the identification of 25 species of which the most representative were Macrophomina phaseolina, Cercospora sesami, Corynespora cassiicola, Alternaria simsimi, Alternaria porri, Fusarium oxysporum, F. fujikuroi, F. equiseti, Colletotrichum capsici, and C. gloesporiodes. The present study showed that diseased sesame plants collected from different production sites in Burkina Faso housed several species of fungi. The fungi presence in diseased plants indicates the need to inform and raise the stakeholders’ awareness about the phytosanitary problems of sesame, but also to develop effective and appropriate control methods against these crop pathogens in Burkina Faso.展开更多
The present experiment was conducted to investigate a dry fish fungus, Cunnighamella blakesleeana, which was identified from the infected part of the Corica soborna, locally named as Kachki fish. Mycelium was hyaline,...The present experiment was conducted to investigate a dry fish fungus, Cunnighamella blakesleeana, which was identified from the infected part of the Corica soborna, locally named as Kachki fish. Mycelium was hyaline, often with granular content, and conidiophores were erected, with verticillate or solitary branches. Zygospores were globose, tuberculate, suspensors equal, smooth, hyaline and heterothallic. Using ITS4 and ITS5 primers, the 740 bp-long ITS region was amplified and sequenced. The ITS region sequences had reciprocal homologies of 98% to 100%. The findings showed that several species of C. blakesleeana fall into the same cluster. It has been determined by molecular data that the fungus we had studied was C. blakesleeana. The maximum mycelial growth (95.33 mm) was observed in the PDA medium, followed by the PSA medium, and the lowest growth (65.50 mm) was measured in the HPA medium in the study of the impact of culture media on the mycelial growth of C. blakesleeana. The influence of temperature on the radial mycelial growth of C. blakesleeana on PDA medium was investigated through five different temperatures. Although pH is a crucial factor in understanding the ecology of spoilage fungus, the highest mycelial growth of C. blakesleeana (88.25 mm) was seen at pH 7, followed by pH 8 and pH 6, while pH 9 was revealed to have the lowest mycelial growth. The outcome suggested that C. blakesleeana thrived in neutral environments.展开更多
Centella asiatica (L.), frequently known as Thankuni, is an important ethnobotanical plant in Bangladesh. This study was conducted to evaluate the morphological characteristics, cultural factors and molecular identifi...Centella asiatica (L.), frequently known as Thankuni, is an important ethnobotanical plant in Bangladesh. This study was conducted to evaluate the morphological characteristics, cultural factors and molecular identification of the causal agent of Alternaria leaf blight disease of C. asiatica. The potato dextrose agar (PDA) medium recorded the maximum mycelial growth (69 mm), followed by the yeast extract agar (YEA) medium, while the honey peptone agar (HPA) medium recorded the lowest growth (27 mm). The optimal pH and temperature for mycelial growth of Alternaria alternata were 6 and 30°C, respectively. Internal transcribed spacer (ITS) region of Alternaria alternata PCR products measured 558 bp and blast search showed 99% sequence similarity with Alternaria alternata species complex. To the best of our knowledge, Alternaria leaf blight disease caused by Alternaria alternata is the first record in Bangladesh.展开更多
This study was conducted in order to determine the fungi and bacteria associated with tomato plants at Cameron Highlands Malaysia. The fungi which have been isolated and detected from tomato plants were: Fusarium oxys...This study was conducted in order to determine the fungi and bacteria associated with tomato plants at Cameron Highlands Malaysia. The fungi which have been isolated and detected from tomato plants were: Fusarium oxysporum, F. solani, F. acuminatum, Rhizoctonia solani, Colletotrichum boninense, C. acutatum and Phoma destructiva. The bacteria which have been isolated and detected from tomato plants were: Ralstonia solanacearum, Xanthomonas vesicatoria, X. gardneri and Pseudomonas syringae. While the most pathogenic fungi were C. boninense, P. destructive and F. oxysporum with the disease incidence (89.6%, 86.6%, 85.6%) respectively, the most pathogenic bacteria were X. vesicatoria and R. solanacearum with the disease incidence (96.6% and 87.6%) respectively.展开更多
Objective:To investigate the prevalence of cercarial infections in freshwater snails from several water sources in Nakhon Nayok,Nonthaburi,and Pathum Thani provinces of Central Thailand,and to reconstruct a phylogenet...Objective:To investigate the prevalence of cercarial infections in freshwater snails from several water sources in Nakhon Nayok,Nonthaburi,and Pathum Thani provinces of Central Thailand,and to reconstruct a phylogenetic tree for improved understanding of the relationships in the cercarial stage.Methods:The snail specimens were collected from 34 total sampling sites and investigated for cercarial infections using the crushing method.The cercarial specimens were classified and used for the phylogenetic tree analysis using the Internal Transcribed Spacer 2(ITS2).Results:A total of 1921 snail specimens were classified into five families and seven species.The results showed that four snail species were identified as intermediate hosts of the larval stages of trematodes,with an overall prevalence of infection of 2.45%(47/1921).The infected snail specimens included five groups of the cercarial type:cercariaeum cercariae,echinostome cercaria,megalurous cercaria,parapleurolophocercous cercaria,and xiphidiocercariae.This is particularly true of xiphidiocercariae,which was found to be the dominant type among cercarial infections in bithyniid snails by approximately 38.00%.With regard to molecular identification,the phylogenetic tree was reconstructed using the neighbor-joining method with 10000 bootstraps and separated the trematodes into three clades:Echinostomatoidea,Microphalloidea and Opisthorchioidea.Conclusions:The study reveals a high prevalence of cercarial infection for each cercarial type and maturation into a definite trematode genus and delineates morphological characteristics and evolutionary trends among each larval trematode in Nakhon Nayok,Nonthaburi and Pathum Thani provinces.In addition,the ITS2 sequence data of cercariae could be used to examine classification of these species at the family level.展开更多
[Objectives]The infection symptoms of cyst nematodes were found in Yuexi, Sichuan. In order to identify the pathogen, the isolated nematodes were identified by morphology and molecular biology. [Methods] The potato ro...[Objectives]The infection symptoms of cyst nematodes were found in Yuexi, Sichuan. In order to identify the pathogen, the isolated nematodes were identified by morphology and molecular biology. [Methods] The potato roots and soil around the roots were collected, the nematodes in the roots were stained and observed, and the cysts were separated by the simple floating method. The second-stage juveniles(J2 s), females and cysts were found, and they were photographed and morphologically measured. The DNA of cysts and J2 s were extracted and identified by species-specific PCR. The DNA sequences of 18 S gene, 28 S D2-D3 region and ITS region in ribosomal DNA were obtained. Sequences of some cyst nematode species were downloaded from GenBank for sequence alignment, and MrBayes 3.2.3 software was used to construct a Bayesian phylogenetic tree. [Results] The nematodes could invade hosts’ root system. The basal knobs of J2 s was nearly round and inclined backward;the average length of the stylet was shorter than 23 μm;the Granek ratio of cysts was greater than or equal to 3, which is highly consistent with that of Globodera rostochiensis. The DNA templates of three cysts and four J2 s were amplified by species-specific PCR, and a band of about 430 bp was obtained. After further sequencing, the length was 434 bp, which is consistent with G. rostochiensis. The evolutionary analysis of rDNA 18 S and 28 S showed that the cyst nematode population was a Globodera species, and further evolutionary analysis of ITS gene confirmed that the population was G. rostochiensis. [Conclusions] The nematodes are G. rostochiensis, which is a quarantine species of great concern to both domestic and import quarantine. Once introduced and colonized, it is hard to eradicate. It is necessary to establish a monitoring system as soon as possible, strengthen the quarantine, supervision and management, increase investment in G. rostochiensis research and development, and develop detection and control technologies, so as to escort the healthy and sustainable development of China’s potato industry.展开更多
Centella asiatica (L.) is commonly known as Thankuni plant and has ethnobotanical importance in Bangladesh. Present experiment was conceded to investigate the wilt disease of C. asiatica, vegetative growth and molecul...Centella asiatica (L.) is commonly known as Thankuni plant and has ethnobotanical importance in Bangladesh. Present experiment was conceded to investigate the wilt disease of C. asiatica, vegetative growth and molecular characterization of pathogenic fungi. Pathogenic fungus, Fusarium equiseti was identified as a causal agent of wilt disease in C. asiatica. The effect of culture media on the mycelial growth of F. equiseti showed the highest (89.25 mm) on potato dextrose agar (PDA) medium followed by carrot agar (CA) medium and the lowest growth (40.25 mm) was measured in HA medium. The optimal temperature and pH for mycelial growth of F. equiseti were 30°C and 7, respectively. The genetic variation of the selected species of fungi, the internal transcribed spacer (ITS) region was amplified using ITS4 and ITS5 primers and sequenced. The PCR product of the ITS region of F. equiseti was 535 bp. Phylogenetic tree of thirty-seven strains of Fusarium sp. based on the nucleotide sequences of the ITS region using the neighbor-joining method with 1000 bootstrapping indicated that 98% - 100% identity with MN886590.1 JUF0046 (F. equiseti). ITS sequences are generally constant, or show little variation within species, but vary between species in a genus. The ITS region is relatively short and can be easily amplified by PCR using universal single primer pairs. Genetic distance exhibited high level of similarity with identical ITS sequences. To date, no published research articles are found on the molecular identification of F. equiseti, the causal agent of fusarium wilt disease of C. asiatica in Bangladesh.展开更多
Objective: To investigate the prevalence of cercarial trematode infection in snails and to examine the reconstruction of the phylogenetic relationship to explain the molecular system of cercarial stage trematodes to e...Objective: To investigate the prevalence of cercarial trematode infection in snails and to examine the reconstruction of the phylogenetic relationship to explain the molecular system of cercarial stage trematodes to estimate the infection rate of in the definite host from the Chao-Phraya Basin.Methods: The snails were collected from 10 provinces of the Chao-Phraya Basin,Thailand by stratified sampling method. The snails were examined for cercarial infection by the crushing method. All DNA specimens were amplified with internal transcribed spacer 3(ITS3) and ITS4 primer based on PCR technique. The sequence data were aligned and used to reconstruct the phylogenetic tree by unweighted pair-group method with arithmetic means with 10 000 bootstraps.Results: The overall rate of cercarial infection was found to be 5.90%(122/2 067). Snails in the family Thiaridae were found to be in the highest prevalence followed by Lymnaeidae, Bithyniidae, Planorbidae, Viviparidae, and Ampullariidae, respectively, while the Buccinidae family(Clea helena) did not reveal any infections. The frequently found species of cercariae were parapleurolophocercous cercariae, cercariae and megarulous cercariae. The monophyletic tree separated the snails into five groups comprised of Heterophyidae, Strigeidae, Lecithodendriidae, Philophthalmidae and Echinostomatidae using the sequence of Angiostrongylus cantonensis as an out-group.Conclusions: This study was the first to report on cercarial infection in the Chao-Phraya Basin, Thailand. This revealed that a high variety of freshwater snails were infected by cercariae stage trematodes with a high prevalence. The sequence data of ITS2 can be used to investigate the phylogenetic relationships of trematodes at the family level and in each clade of different families separated by the definitive hosts.展开更多
The technology based on specific PCR amplification using internal transcribed spacer 2 of nuclear ribosomal DNA for molecular identification and detection of Trichogramma species was studied. Firstly the ITS2s of six ...The technology based on specific PCR amplification using internal transcribed spacer 2 of nuclear ribosomal DNA for molecular identification and detection of Trichogramma species was studied. Firstly the ITS2s of six Trichogramma species were cloned and sequenced, and the interspecific sequence variation was analyzed. Secondly the ITS2 regions of six geographical populations of T. dendrolimi were cloned and sequenced, and the intraspecific sequence identity was analyzed. The results show that the interspecific variation and intraspecific similarity of ITS2 sequences are very suitable for designation of specific primers at specieslevel. Screening of specific primers for T. dendrolimi leads to final sensitive and stable diagnostic primers. This system lets non-specialists can not only identify adults (males and females), but also identify eggs in parasitized hosts rapidly and accurately, which is impossible by conventional methods. Further development of this protocol can create a complete set of specific primers for different species of the whole genus Trichogramma.展开更多
Acrochaete leptochaete (Huber) Nielsen (Chaetophoraceae, Chlorophyta) was isolated from the macroalgae Chaetomorpha collected from intertidal pools in Rongcheng, Shandong, China. 18S rDNA combined with ITS regions wer...Acrochaete leptochaete (Huber) Nielsen (Chaetophoraceae, Chlorophyta) was isolated from the macroalgae Chaetomorpha collected from intertidal pools in Rongcheng, Shandong, China. 18S rDNA combined with ITS regions were used to ascertain the morphological identification of the isolated material. Based on the unialgal culture, asexual reproduction and growth characteristics of A. leptochaete were investigated over wide ranges of temperature and irradiance. Results revealed that asexual reproduction of A. leptochaete could be realized by biflagellate zoospores. The zoospores germinated directly to give self- replicating generations. Zoospore germination was bipolar. A temperature range from 13-21°C and a lower irradiance of 36 μmol/(m2·s) were most favorable for the growth of A. leptochaete. Thallus organization, an important taxonomic criterion for the genus Acrochaete, was affected markedly by temperature and irradiance. Our results extend the knowledge about the species' general biology and its morphological plasticity. For classification and identification of a simple microphytic algae like A. leptochaete, which are traditionally placed in the class Chaetophoraceae, we propose that molecular tools associated with culture observations are applied.展开更多
[Objective] The aim was to study the molecular identification and cultivar fingerprints of Prunus persica (L.) Batsch germplasms.[Method] Sixty peach genotypes,representing China common local cultivars and European sa...[Objective] The aim was to study the molecular identification and cultivar fingerprints of Prunus persica (L.) Batsch germplasms.[Method] Sixty peach genotypes,representing China common local cultivars and European samples were screened by microsatellites (simple sequence repeats,SSRs) and Inter-Simple Sequence Repeat (ISSR) markers.[Result] 26 reproducible bands were amplified by Nine SSR primers,and 24 of which were polymorphic; 236 bands were amplified by 30 ISSR primers,and 113 of which were polymorphic.31 genotypes were discriminated with 1-3 distinct polymorphic bands generated from the primers ISSR and SSR.Seven cultivar-specific ISSR fragments and two SSR unique alleles obtained from this study were available to be converted into Sequence Characterized Amplified Region (SCAR) markers.The genetic similarity coefficient (GS) estimated from these molecular data averaged were 0.939 (ranged from 0.856 to 0.983) for ISSR and 0.646 (ranged from 0.240 to 1.000) for SSR,respectively.The combined grouping association indicated that most local Chinese peach cultivars and exotic accessions were clustered together.This could be related to the mode of introduction and maintenance of the peach cultivars involving limited foundation germplasm,exchange of cultivars between plantations,and periodic development of new recombinant cultivars following sexual reproduction.[Conclusion] The results obtained in this work would help to improve the conservation,molecular identification and management of peach germplasm in breeding.展开更多
Rhizoma Menispermi, derived from the rhizoma of Menispermum dauricum DC., is one of the most popular Chinese medicines. However Rhizoma Menispermi is often illegally mixed with other species in the herbal market, incl...Rhizoma Menispermi, derived from the rhizoma of Menispermum dauricum DC., is one of the most popular Chinese medicines. However Rhizoma Menispermi is often illegally mixed with other species in the herbal market, including Aristolochia mollissimae Hance, which is toxic to the kidneys and potentially carcinogenic. The use of DNA barcoding to authenticate herbs has improved the management and safety of traditional medicines. In this paper, 49 samples belonging to five species, including 34 samples of M. dauricum, from different locations and herb markets in China were collected and identified using DNA barcoding. The sequences of all 34 samples of Rhizoma Menispermi are highly consistent, with only one site variation in internal transcribed spacer 2 (ITS2) of nuclear ribosomal DNA and no variations in the psbA-trnH region. The intra-specific genetic distance is much smaller than inter-specific one. Phylogenetic analysis shows that both sequences allow the successful identification of all species. Nearest distance and BLAST1 methods for the ITS2 and psbA-trnH regions indicate 100% identification efficiency. Our research shows that DNA barcoding can effectively distinguish Rhizoma Menispermi from its adulterants from both commercial and original samples, which provides a new and reliable way to monitor commercial herbs and to manage the modern medicine market.展开更多
We aimed to develop a set of single nucleotide polymorphism(SNP) markers that can be used to distinguish the main cultivated grape(Vitis L.) cultivars in China and provide technical support for domestic grape cultivar...We aimed to develop a set of single nucleotide polymorphism(SNP) markers that can be used to distinguish the main cultivated grape(Vitis L.) cultivars in China and provide technical support for domestic grape cultivar protection, cultivar registration, and market rights protection. A total of 517 high-quality loci were screened from 4 241 729 SNPs obtained by sequencing 304 grape accessions using specific locus amplified fragment sequencing, of which 442 were successfully designed as Kompetitive Allele Specific PCR(KASP) markers. A set of 27 markers that completely distinguishes 304 sequenced grape accessions was determined by using the program, and 26 effective markers were screened based on 23 representative grape cultivars. Finally, a total of 46 out of 48 KASP markers, including 22 markers selected by the research group in the early stage, were re-screened based on 348 grape accessions. Population structure, principal component, and cluster analyses all showed that the 348 grape accessions were best divided into two populations. In addition, cluster analysis subdivided them into six subpopulations. According to genetic distance, V. labrusca, V. davidii, V. heyneana, and V. amurensis were far from V. vinifera, while V. vinifera×V. labrusca and V. amurensis×V. vinifera were somewhere in between these two groups. Furthermore, a core set of 25 KASP markers could distinguish 95.69% of the 348 grape accessions, and the other 21 markers were used as extended markers. Therefore, SNP molecular markers based on KASP typing technology provide a new way for mapping DNA fingerprints in grape cultivars. With high efficiency and accuracy and low cost, this technology is more competitive than other current identification methods. It also has excellent application prospects in the grape distinctness, uniformity, and stability(DUS) test, as well as in promoting market rights protection in the near future.展开更多
Plant diseases generate challenging problems in commercial, agriculture and pose real economic threats to both conventional and organic farming systems. The red palm weevil (Rhynchophorus ferrugineus) (RPW) is one of ...Plant diseases generate challenging problems in commercial, agriculture and pose real economic threats to both conventional and organic farming systems. The red palm weevil (Rhynchophorus ferrugineus) (RPW) is one of the most destructive pests of palms in the world. Nowadays, control methods revolve around treatments based on chemicals, biotechnological systems using semi-chemicals or the development of the sterile insect technique (hardly sustainable at this time) and biological control. Biological control as the use of natural microorganisms, extracted products from microorganisms or genetically improved to resist or eliminate of pathogens. Our aim was to evaluate the entomopathogenicity of indigenous Beauveria bassiana and Metarhizium anisopliae obtained in Gaza strip against larvae and adults of R. ferrugineus in order to identify indigenous strains potentially suitable for Red Palm Weevil biological control. B. bassiana & M. anisopliae were isolated from larvae and adult dead of RPW from different position of Gaza strip. Morphological analysis of the isolated fungi and molecular identification was determined using PCR technique. Also, the efficiency of the isolated fungi were evaluated under lab conditions and optimized as a biological agent. On the anther hand, the ability of treated RPW male to infect females is examined and calculated using Abbott's formula. Our results showed that the B. bassiana and M. anisopliae exhibited a good biological control agent against larvae and adults of RPW. The pathogenicity of the two most virulent isolates and the toxicity assay on larvae showed a highest mortality percentage nearly to 100% by 6 days after spraying the larvae with 3.4 × 108 spores/ml of B. bassiana. The mortality percentage reaches to 90% after spraying the larvae with 3.6 × 108 spores/ml of M. anisopliae. The mortality for the adults treated with pesticide arrives to 50% and the control group 10% at the same time. The results revealed that the infection of the adult males by Entomopathogenic fungi (EPF) can be disseminated into the healthy population, after RPW treatment with B. bassiana and M. anisopliae. Our research concludes that B. bassiana and M. anisopliae locally isolated can be used as biological very effective.展开更多
Flower blight on anthurium(Anthurium andraeanum)was observed during August 2018 on an anthurium cultivation farm in the Songkhla Province of southern Thailand.The fungal isolate was identified as Neopestalotiopsis cla...Flower blight on anthurium(Anthurium andraeanum)was observed during August 2018 on an anthurium cultivation farm in the Songkhla Province of southern Thailand.The fungal isolate was identified as Neopestalotiopsis clavispora based on the morphology and DNA sequence of the internal transcribed spacer(ITS),translation elongation factor 1-α(tef1-α),andβ-tubulin(tub)genes.The phylogenetic tree,based on the combined sequences of ITS,tef1-α,and tub,confirmed this pathogen as N.clavispora.Pathogenicity of the species was confirmed according to Koch’s postulate:N.clavispora could infect anthurium.To the best of our knowledge,this is the first report of N.clavispora as a pathogen of anthurium.展开更多
The systematic classification of the Eucheumatoideae is difficult because of their variable morphology and interpretation of reproductive structures. Kappaphycus and Eucheuma specimens cultivated on the Hainan and Fuj...The systematic classification of the Eucheumatoideae is difficult because of their variable morphology and interpretation of reproductive structures. Kappaphycus and Eucheuma specimens cultivated on the Hainan and Fujian coast of China were introduced from Vietnam, the Philippines and Indonesia. Combined with morphological characteristics, all Kappaphycus and Eucheuma cultivated strains were identified by internal transcribed spacer (ITS) sequences. The phylogenetic tree was constructed using neighbor-joining and maximum likelihood methods. The results indicate that different ITS sequence lengths occurred in the different genera and species. An obvious difference in morphology could be found in the protuberance shape between Kappaphycus and Eucheuma. The protuberance in Eucheuma was thorn-like and in Kappaphycus was wartlike or papillate. Their ITS sequence lengths differed significantly in nucleotide variation rates up to 58.55%-63.90%. All nucleotide variations occurred in the ITS1 and ITS2 regions except for five nucleotide transversions in the 5.8S rDNA region. In addition, the difference was at the branches among congeneric species. Kappaphycus sp. had branches with small buds, while K. alvarezii did not have such a feature. The nucleotide variation rates varied from 7.02% to 7.48% among species; within the same species of the clades it was <1.20%. Eucheumatoideae algae cultivated in China consisted of three clades, K. alvarezii, Kappaphycus sp., and E. denticulatum. The results indicate that ITS sequence analysis was an effective way for identification of interspecies and intraspecies phylogenetic relationships and might provide a clue for molecular identification of algal Eucheumatoideae.展开更多
Death of infants from diarrhoea is a common occurrence in sub-Saharan Africa. This is attributed to unhygienic practices which aid the proliferation of diarrhoea-causing microorganisms. Among these microorganisms, Cam...Death of infants from diarrhoea is a common occurrence in sub-Saharan Africa. This is attributed to unhygienic practices which aid the proliferation of diarrhoea-causing microorganisms. Among these microorganisms, Cam- pylobacter species have been reported as one of the causal agents, Campylobacter spp. are human intestinal pathogens of global importance and their pathogenicity mechanisms are not well understood. This study was designed to investigate the molecular characterisation of Campylobacter gotten from cultural methods in Osun State. Campylobacters isolated were biochemically characterized and biotyped. Confirmation of Campylobacter was done using flaA gene, hippuricase O for Campylobacter jejuni and aspartokinase gene for Campylobacter coli and single locus sequencing glnA gene were performed by PCR. Twenty five samples were amplified by PCR out of 57 Campylobacter strains that were positive for cultural methods from 815 stool samples with diarrhoea and 100 stool samples without diarrhoea. No Campylobacter was isolated from stools of children in the control group. Twenty-five isolates comprising of 18 Campylobater jejuni and 7 C. coli were identified. The nucleotide sequence of the gln A for all the isolated Campylobacter spp. showed 91.0% similarity with the ones in the GenBank. The C. jejuni was classified into biotypes I (44.4%) and II (55.6%) and all C. coli were of biotype I.展开更多
The dried shellfish products with rich nutrients and low-calorie content are favorite food in China,especially in coastal areas.However,the species of dried shellfish products in the market are usually unknown,as the ...The dried shellfish products with rich nutrients and low-calorie content are favorite food in China,especially in coastal areas.However,the species of dried shellfish products in the market are usually unknown,as the taxonomic features were removed during the production process.This study described the application of DNA barcoding technique to the identification of 100 dried shellfish(scallop,squid,octopus and cuttlefish)products in markets.Samples were authenticated by comparing mitochondrial cytochrome oxidase subunit I(COI)gene and 16S ribosomal RNA(16S rRNA)gene sequences with public reference taxonomic databases.The results showed that all the 100 products can be identified at species level.Sixty four scallop adductor products were processed using the bay scallop,Argopecten irradians,and one was from Portuguese oyster,Crassostrea angulata.All the 27 squid,2 cuttlefish and 6 octopus products were produced by the Jumbo flying squid,Dosidicus gigas.The neighbour-joining tree is in agreement with the results of DNA barcoding analysis.The 64 scallop samples formed one A.irradians cluster,leaving Sca65 clustered with the reference oyster sequence C.angulata(MH997922).All the 35 cephalopod(squid,octopus and cuttlefish)samples formed a D.gigas cluster.This investigation revealed a low variety of dried shellfish products sold on the market,and highlighted the high rate of mislabeling and species substitution.Our present work provides a practical method for tracing and authenticating shellfish products.展开更多
文摘Bovine tuberculosis (bTB) is an endemic zoonosis significantly affects animal health in Burkina Faso. The primary causative agent is Mycobacterium tuberculosis (M. tuberculosis) complex, mainly M. bovis. Cattle are considered as natural reservoir of M. bovis. However, in Burkina Faso, the circulation of these strains remains poorly understood and documented. This study aimed to identify and characterize Mycobacterium strains from suspected carcasses during routine meat inspection at Bobo-Dioulasso refrigerated slaughterhouse. A prospective cross-sectional study was conducted from January 2021 to December 2022 on cases of seizures linked to suspected bovine tuberculosis. Microbiological and molecular analyzes were used for mycobacterial strain isolation and characterization. Out of 50 samples, 24% tested positive by microscopy and 12% by culture. Molecular analysis identified 6 strains of Mycobacteria, exclusively Mycobacterium bovis specifically the subspecies bovis (Mycobacterium bovis subsp bovis). In conclusion, M. bovis subsp bovis is the primary agent responsible for bovine tuberculosis in Bobo-Dioulasso. Continuous monitoring of mycobacterial strains is therefore necessary for the effective control of this pathology in the local cattle population.
文摘The current study was performed to evaluate the liver function status as well as molecular characterization of the recovered worms in rats experimentally infected with F. hepatica. Sixteen male Wister rats aged 30 days were randomly allocated into two groups (n = 8). The first group was infected orally with 15 viable encysted metacercaria of F. hepatica per animal. The other group was kept non-infected (control group). At zero time (before infection), the 2<sup>nd</sup>, 4<sup>th</sup>, 6<sup>th</sup>, 8<sup>th</sup>, 10<sup>th</sup>, 12<sup>th</sup> and 14<sup>th</sup> weeks post-infection (WPI), blood and serum samples were collected via puncture of retro-orbital plexus of veins from each rat. Serum enzyme level (AST and ALT) and total protein were measured, and the serum protein profile was carried out using agarose gel electrophoresis. During the period of the experiment, serum ALT and AST activities and serum total globulins significantly increased while serum total proteins and albumin markedly decreased in the infected group. On the 14<sup>th</sup> WPI, the data of the electropherogram showed that globulin fractions (α1-, β- and γ-globulin) levels were significantly increased while α2-globulin was markedly decreased in the infected group. The molecular analysis confirmed the amplification of the ITS1, ITS2 and NDI genes of F. hepatica recovered from the infected liver of rats with amplicon sizes of 630, 510 and 560 bp, respectively. Sequencing of the amplified ITS gene resulted in the determination of 3 strains (PP108836, PP108837, and PP108838). Also, analysis of the ITS2 gene resulted in obtaining 3 isolates under the accession numbers (PP109065, PP109066, and PP109067). In conclusion, fasciolosis in the rat model is suitable for routine experimental infections and caused a pronounced liver dysfunction with discharging of the Fasciola eggs in the faeces and the development of adult stages in the bile ducts. Furthermore, molecular techniques are a sensitive tool for the identification and characterisation of the Fasciola parasite.
文摘Sesame is Burkina Faso’s second essential agricultural export after cotton. It’s consequently a supply of income for producers and foreign exchange for the country. However, sesame production is characterized by low average yields of about 538 kg·ha<sup>-1</sup> at the farmer’s field as compared to the potential yield of the improved varieties (1500 - 2000 kg·ha<sup>-1</sup>). Fungal diseases are some of the major constraints to sesame production in Burkina Faso. The present study contributes to the development of means to control pathogenic fungi of this crop, which are responsible for significant losses. The objective is to identify the fungi associated with diseased sesame plant samples. To this end, 149 samples of diseased sesame plants were collected from different production sites located in three agro-climatic zones of the country. The analysis of the samples according to the blotting paper method, based on the morphological characteristics of the fungi, allowed the identification of 18 genera with prevalence rates from 2.68% to 97.98%. The most frequently identified genera were Macrophomina (97.98%), Cercospora (86.57%), Fusarium (85.23%), Phoma (62.41%) and Colletotrichum (61.07%). The results also showed a variable distribution of fungi according to the agro-climatic zone with the predominance of Macrophomina in all three zones. Molecular identification by DNA sequencing of 120 isolates belonging to the different fungi detected allowed the identification of 25 species of which the most representative were Macrophomina phaseolina, Cercospora sesami, Corynespora cassiicola, Alternaria simsimi, Alternaria porri, Fusarium oxysporum, F. fujikuroi, F. equiseti, Colletotrichum capsici, and C. gloesporiodes. The present study showed that diseased sesame plants collected from different production sites in Burkina Faso housed several species of fungi. The fungi presence in diseased plants indicates the need to inform and raise the stakeholders’ awareness about the phytosanitary problems of sesame, but also to develop effective and appropriate control methods against these crop pathogens in Burkina Faso.
文摘The present experiment was conducted to investigate a dry fish fungus, Cunnighamella blakesleeana, which was identified from the infected part of the Corica soborna, locally named as Kachki fish. Mycelium was hyaline, often with granular content, and conidiophores were erected, with verticillate or solitary branches. Zygospores were globose, tuberculate, suspensors equal, smooth, hyaline and heterothallic. Using ITS4 and ITS5 primers, the 740 bp-long ITS region was amplified and sequenced. The ITS region sequences had reciprocal homologies of 98% to 100%. The findings showed that several species of C. blakesleeana fall into the same cluster. It has been determined by molecular data that the fungus we had studied was C. blakesleeana. The maximum mycelial growth (95.33 mm) was observed in the PDA medium, followed by the PSA medium, and the lowest growth (65.50 mm) was measured in the HPA medium in the study of the impact of culture media on the mycelial growth of C. blakesleeana. The influence of temperature on the radial mycelial growth of C. blakesleeana on PDA medium was investigated through five different temperatures. Although pH is a crucial factor in understanding the ecology of spoilage fungus, the highest mycelial growth of C. blakesleeana (88.25 mm) was seen at pH 7, followed by pH 8 and pH 6, while pH 9 was revealed to have the lowest mycelial growth. The outcome suggested that C. blakesleeana thrived in neutral environments.
文摘Centella asiatica (L.), frequently known as Thankuni, is an important ethnobotanical plant in Bangladesh. This study was conducted to evaluate the morphological characteristics, cultural factors and molecular identification of the causal agent of Alternaria leaf blight disease of C. asiatica. The potato dextrose agar (PDA) medium recorded the maximum mycelial growth (69 mm), followed by the yeast extract agar (YEA) medium, while the honey peptone agar (HPA) medium recorded the lowest growth (27 mm). The optimal pH and temperature for mycelial growth of Alternaria alternata were 6 and 30°C, respectively. Internal transcribed spacer (ITS) region of Alternaria alternata PCR products measured 558 bp and blast search showed 99% sequence similarity with Alternaria alternata species complex. To the best of our knowledge, Alternaria leaf blight disease caused by Alternaria alternata is the first record in Bangladesh.
文摘This study was conducted in order to determine the fungi and bacteria associated with tomato plants at Cameron Highlands Malaysia. The fungi which have been isolated and detected from tomato plants were: Fusarium oxysporum, F. solani, F. acuminatum, Rhizoctonia solani, Colletotrichum boninense, C. acutatum and Phoma destructiva. The bacteria which have been isolated and detected from tomato plants were: Ralstonia solanacearum, Xanthomonas vesicatoria, X. gardneri and Pseudomonas syringae. While the most pathogenic fungi were C. boninense, P. destructive and F. oxysporum with the disease incidence (89.6%, 86.6%, 85.6%) respectively, the most pathogenic bacteria were X. vesicatoria and R. solanacearum with the disease incidence (96.6% and 87.6%) respectively.
基金Srinakharinwirot University,Thailand for providing funding(Project Nos.071/2562,184/2563)
文摘Objective:To investigate the prevalence of cercarial infections in freshwater snails from several water sources in Nakhon Nayok,Nonthaburi,and Pathum Thani provinces of Central Thailand,and to reconstruct a phylogenetic tree for improved understanding of the relationships in the cercarial stage.Methods:The snail specimens were collected from 34 total sampling sites and investigated for cercarial infections using the crushing method.The cercarial specimens were classified and used for the phylogenetic tree analysis using the Internal Transcribed Spacer 2(ITS2).Results:A total of 1921 snail specimens were classified into five families and seven species.The results showed that four snail species were identified as intermediate hosts of the larval stages of trematodes,with an overall prevalence of infection of 2.45%(47/1921).The infected snail specimens included five groups of the cercarial type:cercariaeum cercariae,echinostome cercaria,megalurous cercaria,parapleurolophocercous cercaria,and xiphidiocercariae.This is particularly true of xiphidiocercariae,which was found to be the dominant type among cercarial infections in bithyniid snails by approximately 38.00%.With regard to molecular identification,the phylogenetic tree was reconstructed using the neighbor-joining method with 10000 bootstraps and separated the trematodes into three clades:Echinostomatoidea,Microphalloidea and Opisthorchioidea.Conclusions:The study reveals a high prevalence of cercarial infection for each cercarial type and maturation into a definite trematode genus and delineates morphological characteristics and evolutionary trends among each larval trematode in Nakhon Nayok,Nonthaburi and Pathum Thani provinces.In addition,the ITS2 sequence data of cercariae could be used to examine classification of these species at the family level.
基金Supported by Sichuan Science and Technology Program (2021YFN0009)Science and Technology Project of General Administration of Customs(2020HK161)+1 种基金Key Project of Ningbo Public Welfare Science and Technology Program (2021S024)Technology Development Project of Ningbo Joysun Product Testing Service Company (2020ZS003)。
文摘[Objectives]The infection symptoms of cyst nematodes were found in Yuexi, Sichuan. In order to identify the pathogen, the isolated nematodes were identified by morphology and molecular biology. [Methods] The potato roots and soil around the roots were collected, the nematodes in the roots were stained and observed, and the cysts were separated by the simple floating method. The second-stage juveniles(J2 s), females and cysts were found, and they were photographed and morphologically measured. The DNA of cysts and J2 s were extracted and identified by species-specific PCR. The DNA sequences of 18 S gene, 28 S D2-D3 region and ITS region in ribosomal DNA were obtained. Sequences of some cyst nematode species were downloaded from GenBank for sequence alignment, and MrBayes 3.2.3 software was used to construct a Bayesian phylogenetic tree. [Results] The nematodes could invade hosts’ root system. The basal knobs of J2 s was nearly round and inclined backward;the average length of the stylet was shorter than 23 μm;the Granek ratio of cysts was greater than or equal to 3, which is highly consistent with that of Globodera rostochiensis. The DNA templates of three cysts and four J2 s were amplified by species-specific PCR, and a band of about 430 bp was obtained. After further sequencing, the length was 434 bp, which is consistent with G. rostochiensis. The evolutionary analysis of rDNA 18 S and 28 S showed that the cyst nematode population was a Globodera species, and further evolutionary analysis of ITS gene confirmed that the population was G. rostochiensis. [Conclusions] The nematodes are G. rostochiensis, which is a quarantine species of great concern to both domestic and import quarantine. Once introduced and colonized, it is hard to eradicate. It is necessary to establish a monitoring system as soon as possible, strengthen the quarantine, supervision and management, increase investment in G. rostochiensis research and development, and develop detection and control technologies, so as to escort the healthy and sustainable development of China’s potato industry.
文摘Centella asiatica (L.) is commonly known as Thankuni plant and has ethnobotanical importance in Bangladesh. Present experiment was conceded to investigate the wilt disease of C. asiatica, vegetative growth and molecular characterization of pathogenic fungi. Pathogenic fungus, Fusarium equiseti was identified as a causal agent of wilt disease in C. asiatica. The effect of culture media on the mycelial growth of F. equiseti showed the highest (89.25 mm) on potato dextrose agar (PDA) medium followed by carrot agar (CA) medium and the lowest growth (40.25 mm) was measured in HA medium. The optimal temperature and pH for mycelial growth of F. equiseti were 30°C and 7, respectively. The genetic variation of the selected species of fungi, the internal transcribed spacer (ITS) region was amplified using ITS4 and ITS5 primers and sequenced. The PCR product of the ITS region of F. equiseti was 535 bp. Phylogenetic tree of thirty-seven strains of Fusarium sp. based on the nucleotide sequences of the ITS region using the neighbor-joining method with 1000 bootstrapping indicated that 98% - 100% identity with MN886590.1 JUF0046 (F. equiseti). ITS sequences are generally constant, or show little variation within species, but vary between species in a genus. The ITS region is relatively short and can be easily amplified by PCR using universal single primer pairs. Genetic distance exhibited high level of similarity with identical ITS sequences. To date, no published research articles are found on the molecular identification of F. equiseti, the causal agent of fusarium wilt disease of C. asiatica in Bangladesh.
基金Supported by Faculty of Science,Srinakharinwirot University(Grant No.541/2557)
文摘Objective: To investigate the prevalence of cercarial trematode infection in snails and to examine the reconstruction of the phylogenetic relationship to explain the molecular system of cercarial stage trematodes to estimate the infection rate of in the definite host from the Chao-Phraya Basin.Methods: The snails were collected from 10 provinces of the Chao-Phraya Basin,Thailand by stratified sampling method. The snails were examined for cercarial infection by the crushing method. All DNA specimens were amplified with internal transcribed spacer 3(ITS3) and ITS4 primer based on PCR technique. The sequence data were aligned and used to reconstruct the phylogenetic tree by unweighted pair-group method with arithmetic means with 10 000 bootstraps.Results: The overall rate of cercarial infection was found to be 5.90%(122/2 067). Snails in the family Thiaridae were found to be in the highest prevalence followed by Lymnaeidae, Bithyniidae, Planorbidae, Viviparidae, and Ampullariidae, respectively, while the Buccinidae family(Clea helena) did not reveal any infections. The frequently found species of cercariae were parapleurolophocercous cercariae, cercariae and megarulous cercariae. The monophyletic tree separated the snails into five groups comprised of Heterophyidae, Strigeidae, Lecithodendriidae, Philophthalmidae and Echinostomatidae using the sequence of Angiostrongylus cantonensis as an out-group.Conclusions: This study was the first to report on cercarial infection in the Chao-Phraya Basin, Thailand. This revealed that a high variety of freshwater snails were infected by cercariae stage trematodes with a high prevalence. The sequence data of ITS2 can be used to investigate the phylogenetic relationships of trematodes at the family level and in each clade of different families separated by the definitive hosts.
文摘The technology based on specific PCR amplification using internal transcribed spacer 2 of nuclear ribosomal DNA for molecular identification and detection of Trichogramma species was studied. Firstly the ITS2s of six Trichogramma species were cloned and sequenced, and the interspecific sequence variation was analyzed. Secondly the ITS2 regions of six geographical populations of T. dendrolimi were cloned and sequenced, and the intraspecific sequence identity was analyzed. The results show that the interspecific variation and intraspecific similarity of ITS2 sequences are very suitable for designation of specific primers at specieslevel. Screening of specific primers for T. dendrolimi leads to final sensitive and stable diagnostic primers. This system lets non-specialists can not only identify adults (males and females), but also identify eggs in parasitized hosts rapidly and accurately, which is impossible by conventional methods. Further development of this protocol can create a complete set of specific primers for different species of the whole genus Trichogramma.
基金Supported by the National Natural Science Foundation of China (Nos.31070185, 40876081 and 31093440)the Knowledge Innovation Program of Chinese Academy of Sciences (No. KSCX2-YW-Z-018)+6 种基金the Science and Technology Plan Project of Guangdong Province (No. 2011B031100010)the Team Project of Natural Science Foundation of Guangdong Province(No. S2011030005257)the Talent Introduction Projects of Guangdong Province Universities and Collegesthe Grant of Key Laboratory of Integrated Marine Monitoring and Applied Technologies for Harmful Algal Blooms, S.O.A., MATHAB (No. MATHAB20100301)the Science and Technology Plan Project of Shantou City (No. 2011-162)the Startup Projects (Nos. 09400133 and 09400134)the Grant for Youth Teachers of Shantou University, P. R. China
文摘Acrochaete leptochaete (Huber) Nielsen (Chaetophoraceae, Chlorophyta) was isolated from the macroalgae Chaetomorpha collected from intertidal pools in Rongcheng, Shandong, China. 18S rDNA combined with ITS regions were used to ascertain the morphological identification of the isolated material. Based on the unialgal culture, asexual reproduction and growth characteristics of A. leptochaete were investigated over wide ranges of temperature and irradiance. Results revealed that asexual reproduction of A. leptochaete could be realized by biflagellate zoospores. The zoospores germinated directly to give self- replicating generations. Zoospore germination was bipolar. A temperature range from 13-21°C and a lower irradiance of 36 μmol/(m2·s) were most favorable for the growth of A. leptochaete. Thallus organization, an important taxonomic criterion for the genus Acrochaete, was affected markedly by temperature and irradiance. Our results extend the knowledge about the species' general biology and its morphological plasticity. For classification and identification of a simple microphytic algae like A. leptochaete, which are traditionally placed in the class Chaetophoraceae, we propose that molecular tools associated with culture observations are applied.
基金Supported by National Peach Industrial Technology System (nycytx-31-zs-10 )National Science and Technology Support Program (2008BAD98B03-08)+1 种基金National Peach Commonweal Science (Agriculture) Research Projects (3-37)Chengdu Technology Application and Promotion Program (09YTZD986NC-012)
文摘[Objective] The aim was to study the molecular identification and cultivar fingerprints of Prunus persica (L.) Batsch germplasms.[Method] Sixty peach genotypes,representing China common local cultivars and European samples were screened by microsatellites (simple sequence repeats,SSRs) and Inter-Simple Sequence Repeat (ISSR) markers.[Result] 26 reproducible bands were amplified by Nine SSR primers,and 24 of which were polymorphic; 236 bands were amplified by 30 ISSR primers,and 113 of which were polymorphic.31 genotypes were discriminated with 1-3 distinct polymorphic bands generated from the primers ISSR and SSR.Seven cultivar-specific ISSR fragments and two SSR unique alleles obtained from this study were available to be converted into Sequence Characterized Amplified Region (SCAR) markers.The genetic similarity coefficient (GS) estimated from these molecular data averaged were 0.939 (ranged from 0.856 to 0.983) for ISSR and 0.646 (ranged from 0.240 to 1.000) for SSR,respectively.The combined grouping association indicated that most local Chinese peach cultivars and exotic accessions were clustered together.This could be related to the mode of introduction and maintenance of the peach cultivars involving limited foundation germplasm,exchange of cultivars between plantations,and periodic development of new recombinant cultivars following sexual reproduction.[Conclusion] The results obtained in this work would help to improve the conservation,molecular identification and management of peach germplasm in breeding.
文摘Rhizoma Menispermi, derived from the rhizoma of Menispermum dauricum DC., is one of the most popular Chinese medicines. However Rhizoma Menispermi is often illegally mixed with other species in the herbal market, including Aristolochia mollissimae Hance, which is toxic to the kidneys and potentially carcinogenic. The use of DNA barcoding to authenticate herbs has improved the management and safety of traditional medicines. In this paper, 49 samples belonging to five species, including 34 samples of M. dauricum, from different locations and herb markets in China were collected and identified using DNA barcoding. The sequences of all 34 samples of Rhizoma Menispermi are highly consistent, with only one site variation in internal transcribed spacer 2 (ITS2) of nuclear ribosomal DNA and no variations in the psbA-trnH region. The intra-specific genetic distance is much smaller than inter-specific one. Phylogenetic analysis shows that both sequences allow the successful identification of all species. Nearest distance and BLAST1 methods for the ITS2 and psbA-trnH regions indicate 100% identification efficiency. Our research shows that DNA barcoding can effectively distinguish Rhizoma Menispermi from its adulterants from both commercial and original samples, which provides a new and reliable way to monitor commercial herbs and to manage the modern medicine market.
基金provided by the National Key R&D Program of China(2019YFD1001401)the China Agriculture Research System of MOF and MARA(CARS-29-yc-1)the Agricultural Science and Technology Innovation Program of Chinese Academy of Agricultural Sciences(CAAS-ASTIP-2017-ZFRI)。
文摘We aimed to develop a set of single nucleotide polymorphism(SNP) markers that can be used to distinguish the main cultivated grape(Vitis L.) cultivars in China and provide technical support for domestic grape cultivar protection, cultivar registration, and market rights protection. A total of 517 high-quality loci were screened from 4 241 729 SNPs obtained by sequencing 304 grape accessions using specific locus amplified fragment sequencing, of which 442 were successfully designed as Kompetitive Allele Specific PCR(KASP) markers. A set of 27 markers that completely distinguishes 304 sequenced grape accessions was determined by using the program, and 26 effective markers were screened based on 23 representative grape cultivars. Finally, a total of 46 out of 48 KASP markers, including 22 markers selected by the research group in the early stage, were re-screened based on 348 grape accessions. Population structure, principal component, and cluster analyses all showed that the 348 grape accessions were best divided into two populations. In addition, cluster analysis subdivided them into six subpopulations. According to genetic distance, V. labrusca, V. davidii, V. heyneana, and V. amurensis were far from V. vinifera, while V. vinifera×V. labrusca and V. amurensis×V. vinifera were somewhere in between these two groups. Furthermore, a core set of 25 KASP markers could distinguish 95.69% of the 348 grape accessions, and the other 21 markers were used as extended markers. Therefore, SNP molecular markers based on KASP typing technology provide a new way for mapping DNA fingerprints in grape cultivars. With high efficiency and accuracy and low cost, this technology is more competitive than other current identification methods. It also has excellent application prospects in the grape distinctness, uniformity, and stability(DUS) test, as well as in promoting market rights protection in the near future.
文摘Plant diseases generate challenging problems in commercial, agriculture and pose real economic threats to both conventional and organic farming systems. The red palm weevil (Rhynchophorus ferrugineus) (RPW) is one of the most destructive pests of palms in the world. Nowadays, control methods revolve around treatments based on chemicals, biotechnological systems using semi-chemicals or the development of the sterile insect technique (hardly sustainable at this time) and biological control. Biological control as the use of natural microorganisms, extracted products from microorganisms or genetically improved to resist or eliminate of pathogens. Our aim was to evaluate the entomopathogenicity of indigenous Beauveria bassiana and Metarhizium anisopliae obtained in Gaza strip against larvae and adults of R. ferrugineus in order to identify indigenous strains potentially suitable for Red Palm Weevil biological control. B. bassiana & M. anisopliae were isolated from larvae and adult dead of RPW from different position of Gaza strip. Morphological analysis of the isolated fungi and molecular identification was determined using PCR technique. Also, the efficiency of the isolated fungi were evaluated under lab conditions and optimized as a biological agent. On the anther hand, the ability of treated RPW male to infect females is examined and calculated using Abbott's formula. Our results showed that the B. bassiana and M. anisopliae exhibited a good biological control agent against larvae and adults of RPW. The pathogenicity of the two most virulent isolates and the toxicity assay on larvae showed a highest mortality percentage nearly to 100% by 6 days after spraying the larvae with 3.4 × 108 spores/ml of B. bassiana. The mortality percentage reaches to 90% after spraying the larvae with 3.6 × 108 spores/ml of M. anisopliae. The mortality for the adults treated with pesticide arrives to 50% and the control group 10% at the same time. The results revealed that the infection of the adult males by Entomopathogenic fungi (EPF) can be disseminated into the healthy population, after RPW treatment with B. bassiana and M. anisopliae. Our research concludes that B. bassiana and M. anisopliae locally isolated can be used as biological very effective.
基金supported by Prince of Songkla Universitythe Center of Excellence in Agricultural and Natural Resources Biotechnology(Grant No.CoE-ANRB)phase 3。
文摘Flower blight on anthurium(Anthurium andraeanum)was observed during August 2018 on an anthurium cultivation farm in the Songkhla Province of southern Thailand.The fungal isolate was identified as Neopestalotiopsis clavispora based on the morphology and DNA sequence of the internal transcribed spacer(ITS),translation elongation factor 1-α(tef1-α),andβ-tubulin(tub)genes.The phylogenetic tree,based on the combined sequences of ITS,tef1-α,and tub,confirmed this pathogen as N.clavispora.Pathogenicity of the species was confirmed according to Koch’s postulate:N.clavispora could infect anthurium.To the best of our knowledge,this is the first report of N.clavispora as a pathogen of anthurium.
基金Supported by the Shanghai Pujiang Talent Plan Project(No.05PJ14086)Shanghai Excellent Academic Leaders Project(No.08XD14037)Shanghai Hydrobiology Key Disciplines Funded Projects(No.S30701)
文摘The systematic classification of the Eucheumatoideae is difficult because of their variable morphology and interpretation of reproductive structures. Kappaphycus and Eucheuma specimens cultivated on the Hainan and Fujian coast of China were introduced from Vietnam, the Philippines and Indonesia. Combined with morphological characteristics, all Kappaphycus and Eucheuma cultivated strains were identified by internal transcribed spacer (ITS) sequences. The phylogenetic tree was constructed using neighbor-joining and maximum likelihood methods. The results indicate that different ITS sequence lengths occurred in the different genera and species. An obvious difference in morphology could be found in the protuberance shape between Kappaphycus and Eucheuma. The protuberance in Eucheuma was thorn-like and in Kappaphycus was wartlike or papillate. Their ITS sequence lengths differed significantly in nucleotide variation rates up to 58.55%-63.90%. All nucleotide variations occurred in the ITS1 and ITS2 regions except for five nucleotide transversions in the 5.8S rDNA region. In addition, the difference was at the branches among congeneric species. Kappaphycus sp. had branches with small buds, while K. alvarezii did not have such a feature. The nucleotide variation rates varied from 7.02% to 7.48% among species; within the same species of the clades it was <1.20%. Eucheumatoideae algae cultivated in China consisted of three clades, K. alvarezii, Kappaphycus sp., and E. denticulatum. The results indicate that ITS sequence analysis was an effective way for identification of interspecies and intraspecies phylogenetic relationships and might provide a clue for molecular identification of algal Eucheumatoideae.
文摘Death of infants from diarrhoea is a common occurrence in sub-Saharan Africa. This is attributed to unhygienic practices which aid the proliferation of diarrhoea-causing microorganisms. Among these microorganisms, Cam- pylobacter species have been reported as one of the causal agents, Campylobacter spp. are human intestinal pathogens of global importance and their pathogenicity mechanisms are not well understood. This study was designed to investigate the molecular characterisation of Campylobacter gotten from cultural methods in Osun State. Campylobacters isolated were biochemically characterized and biotyped. Confirmation of Campylobacter was done using flaA gene, hippuricase O for Campylobacter jejuni and aspartokinase gene for Campylobacter coli and single locus sequencing glnA gene were performed by PCR. Twenty five samples were amplified by PCR out of 57 Campylobacter strains that were positive for cultural methods from 815 stool samples with diarrhoea and 100 stool samples without diarrhoea. No Campylobacter was isolated from stools of children in the control group. Twenty-five isolates comprising of 18 Campylobater jejuni and 7 C. coli were identified. The nucleotide sequence of the gln A for all the isolated Campylobacter spp. showed 91.0% similarity with the ones in the GenBank. The C. jejuni was classified into biotypes I (44.4%) and II (55.6%) and all C. coli were of biotype I.
基金This work was supported by research grants from the Fundamental Research Funds for the Central Universities(No.201762014)the National Natural Science Foundation of China(No.31772414)the National Natural Science Foundation of Qingdao City(No.20-3-4-16-nsh).
文摘The dried shellfish products with rich nutrients and low-calorie content are favorite food in China,especially in coastal areas.However,the species of dried shellfish products in the market are usually unknown,as the taxonomic features were removed during the production process.This study described the application of DNA barcoding technique to the identification of 100 dried shellfish(scallop,squid,octopus and cuttlefish)products in markets.Samples were authenticated by comparing mitochondrial cytochrome oxidase subunit I(COI)gene and 16S ribosomal RNA(16S rRNA)gene sequences with public reference taxonomic databases.The results showed that all the 100 products can be identified at species level.Sixty four scallop adductor products were processed using the bay scallop,Argopecten irradians,and one was from Portuguese oyster,Crassostrea angulata.All the 27 squid,2 cuttlefish and 6 octopus products were produced by the Jumbo flying squid,Dosidicus gigas.The neighbour-joining tree is in agreement with the results of DNA barcoding analysis.The 64 scallop samples formed one A.irradians cluster,leaving Sca65 clustered with the reference oyster sequence C.angulata(MH997922).All the 35 cephalopod(squid,octopus and cuttlefish)samples formed a D.gigas cluster.This investigation revealed a low variety of dried shellfish products sold on the market,and highlighted the high rate of mislabeling and species substitution.Our present work provides a practical method for tracing and authenticating shellfish products.
